Signal peptidase SpsB coordinates staphylococcal cell cycle, surface protein septal trafficking and LTA synthesis.

Many cell wall anchored surface proteins of Gram-positive bacteria harbor a highly conserved YSIRK/G-S signal peptide (SPYSIRK+), which deposits surface protein precursors at the cell division septum where they are subsequently anchored to septal peptidoglycan. Previously we identified that LtaS-mediated lipoteichoic acid (LTA) synthesis regulates septal trafficking of YSIRK+ proteins in S. aureus. Interestingly, both LtaS and SPYSIRK+ are cleaved by the signal peptidase SpsB, but the biological implications remain unclear. Here we show that SpsB is required for cleaving SPSpA(YSIRK+) of staphylococcal surface protein A (SpA). Depletion of spsB not only diminished SPSpA processing but also abolished SpA septal localization. The mis-localization is attributed to the cleavage activity of SpsB, as an A37P mutation of SPSpA that disrupted SpsB cleavage also abrogated SpA septal localization. Strikingly, depletion of spsB led to aberrant cell morphology, cell cycle arrest and daughter cell separation defects. Localization studies showed that SpsB predominantly localized at the septum of dividing staphylococcal cells. Finally, we show that SpsB spatially regulates LtaS as spsB depletion enriched LtaS at the septum. Collectively, the data suggest a new dual-mechanism model mediated by SpsB: the abundant YSIRK+ proteins are efficiently processed by septal localized SpsB; SpsB cleaves LtaS at the septum, which spatially regulates LtaS activity contributing to YSIRK+ proteins septal trafficking. The study identifies SpsB as a novel and key regulator orchestrating protein secretion, cell cycle and cell envelope biogenesis.

Genomic profiling of methicillin-sensitive Staphylococcus aureus (MSSA) isolates in Kuwait hospitals.

Background: Staphylococcus aureus is an important pathogen that causes mild to invasive infections in hospitals and the community. Although methicillin-susceptible Staphylococcus aureus (MSSA) isolates continue to cause different infections, there is no data on the genetic backgrounds of the MSSA colonizing or causing infections in Kuwait hospitals. This study aimed to investigate MSSA isolated from patients admitted to Kuwait hospitals for antibiotic resistance and genetic backgrounds to understand their clonal composition. Methods: Consecutive MSSA isolates were collected from single patients during two surveillance periods in 2016 and 2021 in 13 public hospitals. The isolates were characterized using antibiogram, staphylococcal protein A (spa) typing, DNA microarray analysis, and multilocus sequence typing (MLST) using standard protocols. Results: A total of 446 MSSA was cultured from different clinical samples in 2016 (n = 240) and 2021 (n = 206). All isolates were susceptible to vancomycin [minimum inhibitory concentration (MIC) ≤ 2 mg/L], teicoplanin (MIC ≤2 mg/L), linezolid (MIC ≤4 mg/L), ceftaroline (MIC ≤2 mg/L), rifampicin, and mupirocin but were resistant to erythromycin (21.3%), clindamycin (14.0%), gentamicin (3.8%), kanamycin (10.5%), fusidic acid (27.0%), tetracycline (6.9%), trimethoprim (23.1%), and ciprofloxacin (35.2%). Molecular typing identified 155 spa types, dominated by t127 (15.0%), t084 (5.4%), t3841 (5.4%), t267 (2.4%), t442 (2.2%), t091 (2.2%), t021 (2.2%), and t003 (2.2%); 31 clonal complexes (CCs); and 56 sequence types (STs). The majority of the isolates (n = 265; 59.4%) belonged to CC1 (20.6%), CC15 (10.9%), CC22 (5.1%), CC30 (7.6%), CC361 (10.1%), and CC398 (4.7%). Discussion: The MSSA isolates belonged to diverse genetic backgrounds dominated by CC1, CC15, CC22, CC30, CC361, and CC398. The distribution of MSSA clones in 2016 and 2021 showed the stability of these clones over time. The study provides the first comprehensive data on the clonal distribution of MSSA in Kuwait hospitals.

Biofilm Dairy Foods Review: Effect of biofilm production on antimicrobial susceptibility of Staphylococcus aureus bovine mastitis strains from the most prevalent Canadian spa types.

Staphylococcus aureus intramammary infections often leads to clinical and subclinical mastitis in dairy cattle. Prediction of disease evolution and treatment efficacy based on the characteristics of disease-causing strains of S. aureus would significantly improve management of dairy herds. To study the impact of biofilm production and the influence of genetic lineage, we selected S. aureus isolates from the most prevalent Canadian spa types associated with bovine mastitis. Antimicrobial susceptibility in planktonic growth and for bacteria embedded in biofilm was compared. PCR was used to detect the bap gene responsible for atypical biofilm formation. All Canadian spa types from dairy cattle were susceptible to the 8 antimicrobial agents tested. Only strain sa3493 from spa type t267 showed a resistance to pirlimycin. However, bacteria producing larger amounts of biofilms better survived the bactericidal action of antimicrobial agents even when exposed to concentrations 64 folds higher than the minimal inhibitory concentration determined for planktonic cultures. Pirlimycin was more effective on bacteria producing low to moderate levels of biofilm compared with vancomycin or ceftiofur. Antimicrobial agents did not affect the viability of spa types t13401 and t605 that were high biofilm producers. While both these spa types produced high amounts of biofilm, only t605 possessed the bap gene. We also found a close relationship between DIM at sampling and the presence of spa type t605 isolates. These results suggest that detection of S. aureus spa type may help predict the effectiveness of antimicrobial therapy and that some spa types are more likely to be retrieved toward the end of the lactation.

Current Role of Conventional Radiology in Rheumatology.

The correct interpretation of the most common radiological findings in rheumatological diseases can be pivotal for an accurate diagnosis. X rays offer immediacy, worldwide availability, affordability, and the ability to detect subtle bone changes. Numerous scoring systems that have been developed to support clinical practice are based on joint damage that can be observed on X rays. Metabolic, degenerative, infective, and inflammatory processes manifest as distinct alterations in joints and their identification can be challenging. In this review, the authors will discuss the most common radiological findings in hands, hips, and the spine that can serve as valuable diagnostic tools.

Functional Characterisation of Surfactant Protein A as a Novel Prophylactic Means against Oncogenic HPV Infections.

Human papillomavirus (HPV) infection poses a significant health challenge, particularly in low- and middle-income countries (LMIC), where limited healthcare access and awareness hinder vaccine accessibility. To identify alternative HPV targeting interventions, we previously reported on surfactant protein A (SP-A) as a novel molecule capable of recognising HPV16 pseudovirions (HPV16-PsVs) and reducing infection in a murine cervicovaginal HPV challenge model. Building on these findings, our current study aimed to assess SP-A's suitability as a broad-spectrum HPV-targeting molecule and its impact on innate immune responses. We demonstrate SP-A's ability to agglutinate and opsonise multiple oncogenic HPV-PsVs types, enhancing their uptake and clearance by RAW264.7 murine macrophages and THP-1 human-derived immune cells. The SP-A opsonisation of HPV not only led to increased lysosomal accumulation in macrophages and HaCaT keratinocytes but also resulted in a decreased infection of HaCaT cells, which was further decreased when co-cultured with innate immune cells. An analysis of human innate immune cell cytokine profiles revealed a significant inflammatory response upon SP-A exposure, potentially contributing to the overall inhibition of HPV infection. These results highlight the multi-layered impact of SP-A on HPV, innate immune cells and keratinocytes and lay the basis for the development of alternative prophylactic interventions against diverse HPV types.

Safety assessment of the process Fucine Film, based on the Reifenhäuser technology, used to recycle post-consumer PET into food contact materials.

The EFSA Panel on Food Contact Materials, Enzymes and Processing Aids (CEP) assessed the safety of the recycling process Fucine Film (EU register number RECYC322), which uses the Reifenhäuser technology. The input material consists of hot caustic washed and dried poly(ethylene terephthalate) (PET) flakes mainly originating from collected post-consumer PET containers, including no more than 5% PET from non-food consumer applications. The flakes are extruded under vacuum into sheets. The recycled sheets are intended to be used at up to 100% for the manufacture of materials and articles for contact with all types of foodstuffs, excluded drinking water and beverages, for long-term storage at room temperature, with or without hotfill. Based on the limited data available, the Panel concluded that the information submitted to EFSA was inadequate to demonstrate that the recycling process Fucine Film is able to reduce potential unknown contamination of the input PET flakes to a concentration that does not pose a risk to human health.

Impact of spa therapy on physical activity, sleep and heart rate variability among individuals with fibromyalgia: Results of an ancillary study.

Spa therapy is recommended to manage symptoms of fibromyalgia, but the physiological mechanisms underlying this improvement have been poorly studied. In an original study, we explored the effect of a 3-week rheumatology spa treatment for fibromyalgia patients on quality of life and with a symptom severity questionnaire. We present here the results of an ancillary study which explored three secondary criteria using objective measurement methods: diurnal actimetry for physical activity analysis, nocturnal actimetry for sleep analysis and heart rate variability. Eighty-three fibromyalgia patients were randomized to participate in an immediate 3-week rheumatological spa therapy, either a start within 6 weeks after inclusion (interventional group, n = 39) or a delayed, start 6 months after inclusion (control group, n = 44). Patients were asked to wear an actimeter (n = 56) to assess diurnal physical activity and sleep quality and a 24-h Holter ECG (n = 60) to assess nocturnal heart rate variability at baseline, 3 months and 6 months after inclusion. Time spent in sedentary and light physical activity was reduced to ∼30 min at 6 months in the interventional group (P = 0.027). Sleep quality and heart rate variability were not improved. Spa therapy made it possible to reduce sedentary activities in patients' daily life for up to 6 months afterwards, concomitant with the improvement in quality of life, pain and fatigue as highlighted in the original Thermalgi study.

Effectiveness of balneotherapy in reducing pain, disability, and depression in patients with Fibromyalgia syndrome: a systematic review with meta-analysis.

Balneotherapy, using heated natural mineral waters at 36-38 °C, presents a comprehensive treatment approach for Fibromyalgia Syndrome (FMS). This study aims to assess the effect of balneotherapy in reducing pain intensity, disability, and depression in patients with FMS. We want to assess this effect at just four time-points: immediately at the end of the therapy, and at 1, 3, and 6 months of follow-up. Following PRISMA guidelines, we conducted an aggregate data meta-analysis, registered in PROSPERO CRD42023478206, searching PubMed Medline, Science Direct, CINAHL Complete, Scopus, and Web of Science until August 2023 for relevant randomized controlled trials (RCTs) that assess the effect of balneotherapy on pain intensity, disability, and depression in FMS patients. Methodological quality was assessed using the Cochrane methodology, and the pooled effect was calculated using Cohen's standardized mean difference (SMD) and its 95% confidence interval (95% CI) in a random-effects model. Sixteen RCTs were included in the meta-analysis. Balneotherapy is effective in reducing pain intensity (SMD - 1.67; 95% CI -2.18 to -1.16), disability (SMD - 1.1; 95% CI -1.46 to -0.7), and depression (SMD - 0.51; 95% CI -0.93 to -0.9) at the end of the intervention. This effect was maintained at 1, 3, and 6 months for pain intensity and disability. Balneotherapy improves both pain intensity and disability in patients with FMS, providing evidence that its positive effects are sustained for up to 6 months of follow-up. Nevertheless, it is important to note that the improvement in depression varies across different temporal phases.

Could SP-A and SP-D Serum Levels Predict COVID-19 Severity?

Given the various clinical manifestations that characterize Coronavirus Disease 2019 (COVID-19), the scientific community is constantly searching for biomarkers with prognostic value. Surfactant proteins A (SP-A) and D (SP-D) are collectins that play a crucial role in ensuring proper alveolar function and an alteration of their serum levels was reported in several pulmonary diseases characterized by Acute Respiratory Distress Syndrome (ARDS) and pulmonary fibrosis. Considering that such clinical manifestations can also occur during Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection, we wondered if these collectins could act as prognostic markers. In this regard, serum levels of SP-A and SP-D were measured by enzyme immunoassay in patients with SARS-CoV-2 infection (n = 51) at admission (T0) and after seven days (T1) and compared with healthy donors (n = 11). SP-D increased in COVID-19 patients compared to healthy controls during the early phases of infection, while a significant reduction was observed at T1. Stratifying SARS-CoV-2 patients according to disease severity, increased serum SP-D levels were observed in severe compared to mild patients. In light of these results, SP-D, but not SP-A, seems to be an eligible marker of COVID-19 pneumonia, and the early detection of SP-D serum levels could be crucial for preventive clinical management.

Phenotypic and genotypic characterization of Staphylococcus aureus strains isolated from otitis externa: Emergence of CC30-spa t019-SCCmec IV carrying PVL as major genotype.

The increasing emergence of Staphylococcus aureus as the primary causative agent of otitis externa has been noted; however, detailed information regarding the molecular characteristics of these strains in Iran remains scarce. The current study aims to investigate both genotypic and phenotypic attributes of S. aureus strains implicated in ear infections. In the present work, we analyzed 60 S. aureus strains isolated from cases of otitis externa over a period of 45 months. The resistance patterns were determined using disk diffusion and microbroth dilution methods. All S. aureus isolates were confirmed by the nucA polymerase chain reaction assay, and their biofilm production was assessed by a microtiter plate assay. Molecular characterization of the isolates was performed using the staphylococcal cassette chromosome mec, multilocus sequence typing, and staphylococcus protein A typing methods. Overall, the results indicated that 44 out of 60 S. aureus isolates (73.3 %) were methicillin-resistant Staphylococcus aureus. Resistance to mupirocin and vancomycin was observed in 13.3 % and 1.7 % of the tested isolates, respectively. Furthermore, out of the 60 S. aureus isolates, 56 strains (93.4 %) were classified as positive biofilm strains at different levels. Twelve distinct clonal lineages were identified. The vast majority of S. aureus isolates belonged to CC30/ST30-MRSA IV/t019 (41.7 %). Among the 31 strong biofilm producers, the majority (64.5 %) belonged to CC30/ST30-MRSA IV/t019 clone. Biofilm negative isolates belonged to CC22/ST22 (2 isolates), CC8/ST585 (one isolate), and CC8/ST8 (one isolate). Our result revealed that about three-quarters of PVL-positive strains belonged to CC30/ST30. Our data confirmed the presence of MSSA strains among CC30/ST30 and CC22/ST22 isolates. The mupirocin resistant isolates (n = 8) belonged to CC8/ST585-MRSA III/t713 (37.5 %), CC8/ST239-MRSA III/t030 (25 %), CC8/ST8-MRSA IV/t008 (12.5 %), CC8/ST239-MRSA III/t037 (12.5 %), and CC22/ST22-MRSA IV/t790 (12.5 %) lineages. The VRSA strain belonged to the CC8/ST8-MRSA IV/t008 lineage, carrying the vanA determinant. iMLSB phenotypes (n = 14) were distributed across different lineages, including CC30/ST30-MRSA IV/t019 (21.5 %), CC30/ST30-MSSA/t021 (21.5 %), CC22/ST22-MSSA/t005 (14.3 %), CC8/ST239-MRSA III/t030 (14.3 %), CC22/ST22-MSSA/t1869 (7.1 %), CC22/ST22-MRSA IV/t790 (7.1 %), CC8/ST239-MRSA III/t037 (7.1 %), and CC1/ST772-MRSA IV/t10795 (7.1 %). These findings highlight significant genotypic diversity and high biofilm formation among our isolates. The frequent occurrence of the CC/ST30 clone in S. aureus strains isolated from otitis externa reflects the emergence of these lineages as a predominant clone in Iran, posing a significant public health concern.

Balneotherapy and cortisol levels: an updated systematic review and meta-analysis.

The main objective of this review is to examine the impact of balneotherapy on serum and salivary cortisol concentrations. A systematic search was conducted in PubMed, Scopus, Web of Science, PEDro, and Google Scholar. The databases were screened from inception up until April 2024. After screening the scientific literature, 845 articles were retrieved and 17 studies, involving a total of 765 participants, were eventually included in the review. Among them, four were randomized controlled trials, five were non-randomized studies, and eight were pre-post studies with no control group. The evidence gathered in this review indicates a significant short-term reduction in cortisol levels in healthy individuals undergoing balneotherapy, particularly those experiencing high levels of stress. Conversely, in patients with rheumatic conditions (especially if elderly), increases in cortisol levels induced by balneotherapy can act as beneficial hormetic stress, reducing inflammatory mediators and improving pain and functional quality of life. The meta-analysis shows an overall trend of reduction in stress hormone levels, more pronounced in the intervention group undergoing mud-balneotherapy compared to the control group, a finding that, however, does not reach statistical significance (g=-0.11 [95% CI: -0.30; 0.08]; p > 0.05). Current scientific evidence demonstrates that balneotherapy has a positive impact on the regulation of cortisol levels. The regulation of the hypothalamic-pituitary-adrenal axis and the beneficial effects observed on health parameters and quality of life allow mud-balneotherapy to be classified as eustressful stimuli useful in preventing stress-related pathologies in healthy individuals and in alleviating symptoms in patients with chronic conditions. Future research on the topic is advised.

Epigenetic regulation of high light-induced anthocyanin biosynthesis by histone demethylase IBM1 in Arabidopsis.

In plant species, anthocyanin accumulation is specifically regulated by light signaling. Although the CONSTITUTIVELY PHOTOMORPHOGENIC1/SUPPRESSOR OF PHYA-105 (COP1/SPA) complex is known to control anthocyanin biosynthesis in response to light, the precise mechanism underlying this process remains largely unknown. Here, we report that Increase in BONSAI Methylation 1 (IBM1), a JmjC domain-containing histone demethylase, participates in the regulation of light-induced anthocyanin biosynthesis in Arabidopsis. The expression of IBM1 was induced by high light (HL) stress, and loss-of-function mutations in IBM1 led to accelerated anthocyanin accumulation under HL conditions. We further identified that IBM1 is directly associated with SPA1/3/4 chromatin in vivo to establish a hypomethylation status on H3K9 and DNA non-CG at these loci under HL, thereby releasing their expression. Genetic analysis showed that quadruple mutants of IBM1 and SPA1/3/4 resemble spa134 mutants. Overexpression of SPA1 in ibm1 mutants complements the mutant phenotype. Our results elucidate the significance and mechanism of IBM1 histone demethylase in the epigenetic regulation of anthocyanin biosynthesis in Arabidopsis under HL conditions.

Nasal carriage of virulent and multidrug resistant Staphylococcus aureus: a possible comorbidity of COVID-19.

BACKGROUND: Staphylococcus aureus (S. aureus) associated with COVID-19 has not been well documented. This cross-sectional study evaluated the association between nasal S. aureus carriage and COVID-19. METHODS AND RESULTS: Nasopharyngeal samples were collected from 391 participants presenting for COVID-19 test in Lagos, Nigeria, and S. aureus was isolated from the samples. Antimicrobial susceptibility test was done by disc diffusion method. All S. aureus isolates were screened for the presence of mecA, panton-valentine leucocidin (PVL) and toxic shock syndrome toxin (TSST) virulence genes by polymerase chain reaction. Staphylococcal protein A (spa) typing was conducted for all the isolates. Participants with COVID-19 had double the prevalence of S. aureus (42.86%) compared to those who tested negative (20.54%). A significant association was seen between S. aureus nasal carriage and COVID-19 (p = 0.004). Antimicrobial sensitivity results showed resistance to oxacillin (100%), cefoxitin (53%), and vancomycin (98.7%). However, only 41% of the isolates harbored the mecA gene, with SCCmecV being the most common SCCmec type. There was no association between the carriage of virulence genes and COVID-19. A total of 23 Spa types were detected, with t13249 and t095 being the two most common spa types. CONCLUSION: This study examined the association between nasal S. aureus carriage and SARS-COV-2 infection. Further research is required to fully explore the implications of S. aureus co-infection with COVID-19.

Hybrid Illumina-Nanopore assembly improves identification of multilocus sequence types and antimicrobial resistance genes of Staphylococcus aureus isolated from Vermont dairy farms: comparison to Illumina-only and R9.4.1 nanopore-only assemblies.

Antimicrobial resistance (AMR) in Staphylococcus aureus is a pressing public health challenge with significant implications for the dairy industry, encompassing bovine mastitis concerns and potential zoonotic threats. To delve deeper into the resistance mechanisms of S. aureus, this study employed a hybrid whole genome assembly approach that synergized the precision of Illumina with the continuity of Oxford Nanopore. A total of 62 isolates, collected from multiple sources from Vermont dairy farms, were sequenced using the GridION Oxford Nanopore R9.4.1 platform and the Illumina platform, and subsequently processed through our long-read first bioinformatics pipeline. Our analyses showcased the hybrid-assembled genome's superior completeness compared to Oxford Nanopore (R9.4.1)-only or Illumina-only assembled genomes. Furthermore, the hybrid assembly accurately determined multilocus sequence typing (MLST) strain types across all isolates. The comprehensive probe for antibiotic resistance genes (ARGs) using databases like CARD, Resfinder, and MEGARES 2.0 characterized AMR in S. aureus isolates from Vermont dairy farms, and revealed the presence of notable resistance genes, including beta-lactam genes blaZ, blaI, and blaR. In conclusion, the hybrid assembly approach emerged as a tool for uncovering the genomic nuances of S. aureus isolates collected from multiple sources on dairy farms. Our findings offer a pathway for detecting AMR gene prevalence and shaping AMR management strategies crucial for safeguarding human and animal health.

Nasal Staphylococcus aureus Carriage and Antimicrobial Resistance Profiles Among Community-Dwelling Adults in Jiangsu, China.

INTRODUCTION: Persistent nasal carriage has been associated with Staphylococcus aureus infection. Previous S. aureus studies in Asia have primarily focused on clinical patients, providing limited information on persistent nasal carriage among the general adult population. METHODS: This study examined 143 healthy adults in a community in Jiangsu, China. Nasal swab samples were collected 10 times. The colonization status was identified using SPA typing. We also determined antimicrobial susceptibility, genotype, and genomic characteristics of S. aureus. RESULTS: The prevalence of S. aureus nasal carriage among the community individuals was on average 16.78%. The carriage rates of methicillin-resistant S. aureus and multidrug-resistant S. aureus were 6.29% and 7.69%, respectively. We identified 8.39% persistent carriers, 39.16% intermittent carriers, and 52.45% noncarriers. Furthermore, family members displayed concordance in terms of genotype and genomic characteristics. CONCLUSION: Persistent nasal sampling captured intermittent carriers that were missed during short-term sampling, thus highlighting the necessity for regular community testing. SPA typing can serve as a rapid method for determining S. aureus colonization. The potential for intrafamilial transmission of S. aureus is evident, with persistent carriers being the most probable source of infection.

Molecular and epidemiological characterization of Staphylococcus aureus causing bovine mastitis in China.

BACKGROUND: Staphylococcus aureus is one of the most prevalent pathogens in bovine mastitis, which leads to substantial financial losses for the dairy industry. RESULTS: In this study, S. aureus (n = 72) was isolated from 18 dairy farms in 15 provinces across China in 2021. The identification of these isolates at the species level was achieved by employing 16S rRNA sequencing. An isothermal amplification method for auxiliary detection of S. aureus was established, which can be employed not only for laboratory detection but also for point-of-care testing (POCT). Molecular characteristics of S. aureus mastitis in Chinese dairy cows were determined through MLST and spa typing. Finally, methicillin-resistant Staphylococcus aureus (MRSA) and MRSA resistance genes were detected using MIC and PCR amplification techniques. 72 isolates were identified as 12 sequence types (STs) and 7 clonal complexes (CC). ST1/CC1 was the dominant prevalent accounting for 33.3 % of the total, and exhibiting a wide distribution range. In terms of spa types, t114 was the dominant type, accounting for 31.9 % of the total, followed by t529 as the second major type. Four S. aureus strains were classified as MRSA according to their levels of oxacillin resistance (MIC ≥4 µg/mL). Among these four MRSA strains, one of them was found to be mecA positive. However, the presence of drug-resistance genes mecA and mecC was not detected in the remaining three MRSA strains, indicating the possible existence of new resistance genes. CONCLUSIONS: Our study investigated the prevalence of S. aureus mastitis in dairy cows in China, while also examined the molecular characteristics and MRSA strains. This information will help with the clinical monitoring, prevention, and control of S. aureus mastitis in dairy cattle.

The Long-Acting Serine Protease Inhibitor mPEG-SPA-MDSPI16 Alleviates LPS-Induced Acute Lung Injury.

Anti-inflammatory drugs have become the second-largest class of common drugs after anti-infective drugs in animal clinical care worldwide and are often combined with other drugs to treat fever and viral diseases caused by various factors. In our previous study, a novel serine protease inhibitor-encoding gene (MDSPI16) with improved anti-inflammatory activity was selected from a constructed suppressive subducted hybridization library of housefly larvae. This protein could easily induce an immune response in animals and had a short half-life, which limited its wide application in the clinic. Thus, in this study, mPEG-succinimidyl propionate (mPEG-SPA, Mw = 5 kDa) was used to molecularly modify the MDSPI16 protein, and the modified product mPEG-SPA-MDSPI16, which strongly inhibited elastase production, was purified. It had good stability and safety, low immunogenicity, and a long half-life, and the IC50 for elastase was 86 nM. mPEG-SPA-MDSPI16 effectively inhibited the expression of neutrophil elastase and decreased ROS levels. Moreover, mPEG-SPA-MDSPI16 exerted anti-inflammatory effects by inhibiting activation of the NF-κB signaling pathway and the MAPK signaling pathway in neutrophils. It also exerted therapeutic effects on a lipopolysaccharide (LPS)-induced acute lung injury (ALI) mouse model. In summary, mPEG-SPA-MDSPI16 is a novel anti-inflammatory protein modified with PEG that has the advantages of safety, nontoxicity, improved stability, and strong anti-inflammatory activity in vivo and in vitro and is expected to become an effective anti-inflammatory drug.

Characterization of PIF4 Phosphorylation by SPA1.

The PHYTOCHROME INTERACTING FACTORs (PIFs) play pivotal roles in regulating thermo- and photo-morphogenesis in Arabidopsis. One of the main hubs in thermomorphogenesis is PIF4, which regulates plant development under high ambient temperature along with other PIFs. PIF4 enhances its own transcription and PIF4 protein is stabilized under high ambient temperature. However, the mechanisms of thermo-stabilization of PIF4 are less understood. Recently, it was shown that SUPPRESSOR OF PHYA-105 1 (SPA1) can function as a serine/threonine kinase to phosphorylate PIF4 in vitro, and the phosphorylated form of PIF4 is more stable under high ambient temperature conditions. In this chapter, we describe the in vitro kinase assay of PIF4 by SPA1. In principle, this protocol can be applied for other putative substrates and kinases.

Measuring moral distress in health professionals using the MMD-HP-SPA scale.

BACKGROUND: Moral distress (MD) is the psychological damage caused when people are forced to witness or carry out actions which go against their fundamental moral values. The main objective was to evaluate the prevalence and predictive factors associated with MD among health professionals during the pandemic and to determine its causes. METHODS: A regional, observational and cross-sectional study in a sample of 566 professionals from the Public Health Service of Andalusia (68.7% female; 66.9% physicians) who completed the MMD-HP-SPA scale to determine the level of MD (0-432 points). Five dimensions were used: i) Health care; ii) Therapeutic obstinacy-futility, iii) Interpersonal relations of the Healthcare Team, iv) External pressure; v) Covering up of medical malpractice. RESULTS: The mean level of MD was 127.3 (SD=66.7; 95% CI 121.8-132.8), being higher in female (135 vs. 110.3; p<0.01), in nursing professionals (137.8 vs. 122; p<0.01) and in the community setting (136.2 vs. 118.3; p<0.001), with these variables showing statistical significance in the multiple linear regression model (p<0.001; r2=0.052). With similar results, the multiple logistic regression model showed being female was a higher risk factor (OR=2.27; 95% CI 1.5-3.4; p<0.001). 70% of the sources of MD belonged to the dimension "Health Care" and the cause "Having to attend to more patients than I can safely attend to" obtained the highest average value (Mean=9.8; SD=4.9). CONCLUSIONS: Female, nursing professionals, and those from the community setting presented a higher risk of MD. The healthcare model needs to implement an ethical approach to public health issues to alleviate MD among its professionals.